Vertebrate Zoology 62 (3) 2012 387

387 – 397 © Museum für Tierkunde Dresden, ISSN 1864-5755, 19.12.2012

Betta siamorientalis, a new species of bubble-nest building fighting

fish (Teleostei: Osphronemidae) from eastern Thailand

Chanon Kowasupat 1, Bhinyo Panijpan 2 *, Pintip Ruenwongsa 2 &

Tadsanai Jeenthong 1, 3
1 Institute for Innovative Learning, Mahidol University, 999 Phuttamonthon 4 Road, Salaya,

Nakhon Pathom 73170, Thailand.

chanonnon@hotmail.com
2 Faculty of Science, Mahidol University, 272 Rama 6 Road, Phayathai, Bangkok 10400, Thailand.
bhinyop@gmail.com, pruenwong@hotmail.com
3
Thailand Natural History Museum, Pathum Thani 12120, Thailand.
tadsanai_j@hotmail.com
* Author to whom correspondence should be addressed.
bhinyop@gmail.com
Accepted on November 05, 2012.
Published online at www.vertebrate-zoology.de on December 10, 2012.

> Abstract
Betta siamorientalis sp. n., a new species of fighting fish belonging to the Betta splendens group, inhabiting four provinces in
the eastern Thailand, is described. The combination of slender body shape with reddish vertical bars on the black opercle and
an apparent black body distinguishes the new species from other congeners especially B. imbellis. The blacker appearance
relative to B. imbellis is due to a higher proportion of black body area relative to that occupied by blue/green iridescence of
the scales. Based on morphological similarity and fin colour pattern and vertical bar colour on the opercle, the new species
appears to be more closely related to B. imbellis and B. splendens than other members of the B. splendens group.

> Key words

Betta siamorientalis sp. n., Black imbellis, Bubble-nesting fighting fish, Thailand.

Introduction

Based on behaviourial and morphological charac-
ters and details of head and body patterns, Witte &
Schmidt (1992) have classified betta fighting fishes
into 15 species groups (see also Tan & Ng, 2005),
one of which is the Betta splendens group. The term
species group is used here for an assemblage of spe-
cies sharing diagnostic characters. Members of the B.
splendens group are bubble-nest building bettas with
the following characters: small head, often brightly
coloured body, iris of eye with iridescent green or
blue patches, body slender, head with parallel opercles
when viewed dorsally, red or brown anal and caudal
fin rays contrasting with iridescent interradial mem-
brane (especially in male) (Witte & Schmidt, 1992).
These members are B. splendens Regan, 1910; B.
smaragdina Ladiges, 1972; B. imbellis Ladiges,1975;
B. stiktos Tan & Ng, 2005; and the newly identified
B. mahachaiensis Kowasupat, Panijpan, Ruenwongsa
& Sriwattanarothai, 2012). It is well recognised that
the native habitats for wild B. smaragdina are in the
northeast and Laos PDR, those for B. imbellis are in
south of Thailand extending to peninsular Malaysia.
Betta splendens is distributed more or less country-
wide from the north to central to the upper southern
peninsula and also from east to west of Thailand and
beyond. Betta mahachaiensis has been found con-
fined to the Samut Sakhon province, west of Bangkok
(Kowasupat et al.,2012). Since recorded history,
388 Kowasupat et al.: Betta siamorientalis, a new fighting fish from eastern Thailand
Thais have reared these fighting fish and bred them,
especially, B. splendens, for both fighting and orna-
mental purposes (Smith, 1927, 1945; Monvises et al.,
2009). However, there are only a few reports on the
distribution of the bubble-nest building fighting fish
in Thailand or nearby countries. For example, Lert­
panich & Aranyavalai (2007) had identified the fish
from the eastern Thailand all as B. splendens based
mainly on the presence of reddish bars on the opercle.
Our ongoing extensive fieldwork since 2007 on Betta
distribution all over Thailand has now covered sixty-
seven provinces (as of 2012) of the total seventy-seven
including all seven provinces in eastern Thailand and
some bordering Cambodia and Laos PDR. It has been
noted by us and others that the betta fighting fish in the
Chachoengsao province and some other eastern prov-
inces possess different characteristics to other known
members of the B. splendens group (authors′ personal
communication). Locals say such unique characteris-
tics have been known for generations, but the red bars
on the opercle have persuaded them to believe the fish
to be B. splendens. This fish has been called black im-
bellis by those familiar with it due to its black opercle
and apparently black body. This black imbellis has
a general appearance quite similar to the fish caught
from Cambodia in Seam Reap by Kühne (2008), who
named it Betta cf. imbellis, and similar to the Betta sp.
in the Priay Khmang village by Linke (2009). Schäfer
(2009) also received from Vietnam some traded fish
and identified it as Betta cf. stiktos based on preserved
specimens of the fish for comparison. However, upon
later comparative work with live wild-caught speci-
mens from the type-locality, he then changed the name
to Betta cf. imbellis and called it Vietnam black em-
phasising the paucity of blue/green iridescence on the
black body.
Upon in-depth analysis, our data suggested that
the fish was a new species, at least those found in
four provinces of eastern Thailand: Chachoengsao,
Sa Kaeo, Prachin Buri and Chonburi (near border of
Chachoengsao) and also at one site in Cambodia across
the border from Sa Kaeo (Kowasupat & Panijpan,
2012; Linke 2012). The aim of this paper is to formally
describe this so-called black imbellis.
Materials and methods
Fighting fish specimens were preserved in 70% etha-
nol for long-term storage. The left side of all speci-
mens was used for morphometric measurements point
to point using digital Vernier caliper readings to the
nearest 0.1 mm according to the method of Schindler
& Schmidt (2006) and Tan & Ng (2005), both modi-
fied from Witte & Schmidt (1992). Ratios are given
in percent standard length or percent head length. The
terminology and general format of description is based
on Witte & Schmidt (1992), Tan & Kottelat (1998)
and Tan & Ng (2005). Meristic counts were carried
out as described by Witte & Schmidt (1992) except
for the number of predorsal scales, which were count-
ed continuously following Ng & Kottelat (1994).
The phylogenetic species concept is used in this study
(Cracraft, 1989; see Tan & Ng, 2005).
All statistical analyses were performed using SPSS
Version 20. To avoid undesirable effects of using ratios
in statistical analyses, the logarithms of these ratios
(LGRAT) were used (Atchley & Anderson, 1978).
The LGRATs of each trait of each species were tested
for normality using the Shapiro-Wilk test. For non-
normal traits, the Kruskal-Wallis one-way ANOVA
together with Mann-Whitney U test were used to
compare means. Otherwise, the Levene′s test was
used to test for homogeneity of variances. If the vari-
ances were equal, the one-way ANOVA together with
the Tukey post-hoc test were used to compare means.
Conversely, the Welch one-way ANOVA and the
Games-Howell post-hoc test were used. Significant
level was adjusted by the Bonferroni correction.
Specimens examined have been deposited at the
Thailand Natural History Museum, Pathum Thani
(THNHM) and also kept in the authors’ own collection
for the Thailand Betta Project (TBP). Abbreviations
used are SL – standard length, HL – head length, and
TL – total length.
Comparative material
The following materials were compared for morpho-
metric and meristic characteristics as well as general
appearance and body shape.
Betta imbellis caught from thirteen provinces of
southern Thailand, one province of Malaysia (type lo-
cality), and one province of Vietnam
(THNHM-F-01557 – 61, THNHM-F-01564 – 6,
THNHM-F-01569, THNHM-F-01571,
THNHM-F-01573, THNHM-F-01575 – 7,
THNHM-F-01579, THNHM-F-01582,
THNHM-F-01585 – 7, THNHM-F-01596 – 600,
THNHM-F-01829, THNHM-F-01841)
Betta splendens caught from thirteen provinces of
central, northern, and western Thailand
(THNHM-F-01673, THNHM-F-01676 – 8,
THNHM-F-01680 – 1, THNHM-F-01684,
THNHM-F-01710, THNHM-F-01712 – 3,
THNHM-F-01716, THNHM-F-01718,
THNHM-F-01719, THNHM-F-01691 – 2,
THNHM-F-01696, THNHM-F-01698 – 704,
THNHM-F-01707 – 8, THNHM-F-01691 – 2).
Vertebrate Zoology n 62 (3) 2012
Results
Betta siamorientalis sp. n.
Figs 1 – 3; Tables 1,2
Holotype. THNHM – F – 01540 (36.1 mm SL), Thailand, Cha­
cho­engsao province, Bang Khla district, Tha Thonglang subdis-
trict, Sai Hai village (13°42′ N, 101°13′ E); coll. C. Kowasupat
et al., 12 Oct. 2010.
Paratypes. THNHM – F – 01541, 10 ex., same locality as ho­lo­
type; coll. C. Kowasupat et al., 12 Oct. 2010. – THNHM-F-01836,
2 ex., same locality as holotype; coll. B. Panijp­ an et al., 1 May
2012. – THNHM-F-01835, 1 ex., same loc­ality as holotype;
coll. C. Kowasupat et al., 8 May 2012. – THNHM-F-01535,
3 ex., Thailand, Chachoengsao province, Bang Khla dis-
trict, Sa Med Nuea subdistrict, behind central aquarist mar-
ket (13°40′ N, 101°13′ E); C. Kowasupat et al. 6 Oct. 2010 –
THNHM-F-01539, 12 ex., Thailand, Chachoengsao province,
Bang Khla district, Sa Med Nuea subdistrict, behind central aq-
uarist market (13°40′ N, 101°13′ E); T. Jeenthong et al. 12 Oct.
2010 – THNHM-F-01538, 10 ex., Thailand, Cha­cho­engsao
province, Bang Khla district, Sa Med Tai subdistrict, Nong Sano
village, near border of Wang Klon village (13°38′ N, 101°11′
E); coll. T. Jeenthong et al. 12 Oct. 2010. –THNHM-F-01823,
5 ex., Thailand, Chachoengsao province, Bang Khla district, Sa
Med Tai subdistrict, Wang Klon village (13°39′ N, 101°10′ E);
coll. T. Jeenthong et al. 20 Dec. 2011. –THNHM-F-01831, 5
ex., Thailand, Chachoengsao province, Bang Khla district, Sa
Med Tai subdistrict, Wang Klon village (13°39′ N, 101°10′ E);
coll. T. Jeenthong et al. 8 May 2012. – THNHM-F-01824, 10
ex., Thailand, Chachoengsao province, Bang Khla district, Sa
Med Tai subdistrict, Nong Sano village, Klum Ban Mor Taek
(13°37′ N, 101°12′ E); coll. B. Panijpan et al. 26 Dec. 2011. –
THNHM-F-01833, 2 ex., Thailand, Chachoengsao province,
Bang Khla district, Sa Med Tai subdistrict, Nong Sano vil-
lage (13°37′ N, 101°12′ E); coll. P. Ruenwongsa et al. 8 May
2012. – THNHM-F-01834, 1 ex., Thailand, Chachoengsao
pro­vince, Bang Khla district (13°37′ N, 101°12′ E); coll. P.
Ruenw ­ ongsa et al. 10 Jun. 2012. – THNHM-F-01825, 8 ex.,
Thai­land, Chachoengsao province, Plaeng Yao district, Ban Bor
sub­­district (13°35′ N, 101°15′ E); coll. P. Ruenwongsa et al. 3
Jan. 2012. – THNHM-F-01836, 1 ex., Thailand, Chachoengsao
province, Phanom Sarakham district, Ko Khanun (Area
1) (13°42′ N, 101°26′ E); coll. A. Sakthawornlert 1 Oct.
2012. – THNHM-F-01837, 1 ex., Thailand, Chachoengsao
province, Phanom Sarakham district, Ko Khanun (Area 2)
(13°41′ N, 101°26′ E); coll. A. Sakthawornlert 1 Oct. 2012. –
THNHM-F-01826, 5 ex., Thailand, Chon Buri province, Phanat
Nik­hom district, pond beside Kok Plor subdistrict administra-
tion building (13°34′ N, 101°8′ E); coll. C. Kowasupat et al.
10 Jan. 2012. – THNHM-F-01827, 3 ex., Thailand, Chon Buri
province, Phanat Nikhom district, Kok Plor subdistrict, Noen
Put village (13°30′ N, 101°6′ E); coll. C. Kowasupat et al. 15
Jan. 2012. – THNHM-F-01531, 2 ex., Thailand, Sa Kaeo prov-
ince, Watthana Nakhon district, Chong Kum subdistrict, Chong
Kum village (Area 1) (13°53′ N, 102°27′ E); coll. C. Kowasupat
et al. 5 Oct. 2010. – THNHM-F-01532, 2 ex., Thailand, Sa
Kaeo province, Watthana Nakhon district, Chong Kum sub-
district, Chong Kum village (Area 2) (13°53′ N, 102°27′ E);
coll. C. Kowasupat et al. 5 Oct. 2010. – THNHM-F-01533,
2 ex., Thailand, Sa Kaeo province, Watthana Nakhon dis-
trict, Chong Kum subdistrict, Chong Klum village (Area 3)
(13°54′ N, 102°28′ E); coll. C. Kowasupat et al. 5 Oct. 2010. –
THNHM-F-01534, 2 ex., Thailand, Sa Kaeo province, Watthana
Nakhon district, Chong Kum subdistrict, Chong Klum village
389
(Area 4) (13°54′ N, 102°28′ E); coll. T. Jeenthong et al. 5 Oct.
2010. – THNHM-F-01542, 3 ex., Thailand, Prachin Buri prov-
ince, Kabin Buri district, Kabin Buri subdistrict, Tung Faek vil-
lage (13°58′ N, 101°42′ E); coll. C. Kowasupat et al. 10 Dec.
2010. – THNHM-F-01543, 2 ex., Thailand, Prachin Buri prov-
ince, Kabin Buri district, Hat Nang Kaeo subdistrict, Bueng
Laem Hin (14°0′ N, 101°39′ E); coll. C. Kowasupat et al. 10
Dec. 2010. – THNHM-F-01544, 3 ex., Cambodia, Serei Sao­
phoan, Banteay Meanchey province, Bod Tong village (13°39′
N, 102°45′ E); coll. B. Panijpan et al. 12 Jun. 2011.
Diagnosis. Betta siamorientalis sp. n. is distinguished
from other members of the B. splendens group by hav-
ing the dark-brown to black body; black opercle with
two parallel reddish vertical bars; red patches on dark-
brown to black opercular membrane; caudal fin rays
with distal red crescent and thin black edge; absence
of dark transverse bars on caudal fin; distal half of pos-
terior anal fin rays red with a small red patch at distal
tip; black and red from proximal pelvic fin base to the
white tip.
Description. General body shape, appearance and col-
our pattern are illustrated in Figs. 1 – 3. Head small,
short (head length 28.0 – 34.0 %SL), eyes large (orbit
diameter 23.7 – 31.6 %HL); body slender (body depth
at dorsal-fin origin 23.6 – 32.1 %SL); opercles parallel
when head viewed dorsally; dorsal fin positioned after
mid-body (predorsal length 57.9 – 67.1 %SL); caudal
fin posterior margin rounded; dorsal fin distal margin
rounded; pelvic fins falcate; anal fin pointed and most
of male distal tip extended to half or more of caudal fin
length (30 out of total 40). Meristic and morphomet-
ric data are summarised in Tables 1 and 2. Maximum
known size is 32.7 mm SL (THNHM – F – 01825).
Colouration. See Fig. 2 for colouration in live speci-
mens. Male with dark-brown to black head, with iri-
descent yellowish-green to bluish-green scales; oper-
cles with double parallel reddish vertical bars (a verti-
cal bar on the edge of opercle redder than the inner one
nearer to the base); opercular membrane dark-brown to
black with red patches; eyes with iridescent yellowish-
green to bluish-green patches at bottom and posterior
regions; body background dark-brown to black with
the iridescent yellowish-green to bluish-green scales:
high ratio (greater than 60%) of body background
(dark-brown to black) relative to scale iridescence
(yellowish-green to bluish-green) in most specimens
making it appear dark; dorsal fin rays dark-brown to
black contrasting with iridescent yellowish-green to
bluish-green interradial membranes, and at least prox-
imal two-thirds having black transverse bars; caudal
and anal fin rays red-brown to black contrasting with
iridescent yellowish-green to bluish-green interradial
membranes; caudal fin with distal red crescent and
thin black edge; caudal fin interradial membranes with
390 Kowasupat et al.: Betta siamorientalis, a new fighting fish from eastern Thailand

Fig. 1. Betta siamorientalis sp. n., THNHM-F-01540, male, holotype, 36.1 mm SL, GenBank Accession Numbers: JQ818630
(COI), JQ815715 (ITS1).

Fig. 2. Betta siamorientalis sp. n., THNHM-F-01823, live adult male displaying aggression, paratype, 28.1 mm SL.

small black spots in some specimens; posterior anal fin
with a small red patch at distal tip (some rays have red
colour near the end, especially, those near the distal
tip); anal fin interradial membranes with small black
dots in some specimens; red-brown to black and red
from proximal pelvic fin base to the white tip; pectoral
fins hyaline. Female less colourful (see Fig. 3): head
and body yellowish to light-brown with iridescent
yellowish-green to bluish-green scales; opercles pale
gold with two faint reddish vertical bars; throat with
chin-bar; body with distinct upper central stripe nearly
parallel to central stripe, at level of eye; caudal-pedun-
cle black spot present; dorsal, caudal, anal, and pelvic
fin rays yellowish with faint iridescent green to bluish-
green interradial membranes; caudal fin with distal
pale red crescent margin and pale thin black edge, or
yellowish on both margin and edge; distal half of pos-
terior anal fin with a small pale red or yellowish patch
at distal tip; other characters similar to male.
Comparative notes. Betta siamorientalis sp. n. is dis­
tinguished from other members of the B. splendens
group by having area belonging to the dark-brown to
black body (background) greater than that of the iri-
descence of the scales (vs. red-brown to black body
background in B. splendens; vs. area belonging to
the dark-brown to black body (background) less than
that of the iridescence of the scales in B. imbellis);
black opercles with two parallel reddish vertical bars
(vs. double iridescent green to bluish-green vertical
bars in B. imbellis and B. mahachaiensis; vs. opercle
with iridescent green scales in B. smaragdina and
B. stiktos); red patches on brown-to-black opercular
membrane (vs. absence in B. mahachaiensis); caudal
Vertebrate Zoology n 62 (3) 2012
Fig. 3. (a) Betta siamorientalis sp. n., THNHM-F-01824, live male (acclimatized), paratype, 30.9 mm SL; (b) Betta siamorientalis
sp. n., THNHM-F-01824, live female (acclimatized), paratype, 26.7 mm SL.
fin with distal red crescent and thin black edge (vs.
absence in B. smaragdina, B. stiktos, and B. maha­
chaiensis); absence of dark transverse bars on caudal
fin (vs. presence in B. stiktos); posterior anal fin rays
red with a small red patch at distal tip (vs. absence in
B. smaragdina, B. stiktos, and B. mahachaiensis); red-
brown to black and red from proximal pelvic fin base
to the white tip (vs. absence in B. stiktos, and B. ma­
hachaiensis); absence of iridescent bluish-green front
margin of pelvic fin (vs. presence in B. stiktos, and B.
mahachaiensis).
Distribution. Based on our extensive survey of fight-
ing fish in sixty-seven (total of seventy-seven as of
2012) provinces, the wild B. siamorientalis sp. n. is
currently found in only 4 provinces in Thailand: Cha­
choengsao, Sa Kaeo, Prachin Buri, and Chon Buri
391
b
(only in areas close to Chachoengsao). The eastern
region of Thailand harbouring this species is shown
in Fig. 4. Betta siamorientalis sp. n. is also found in
Banteay Meanchey, a province of Cambodia on the
eastern border of Sa Kaeo province: this may indi-
cate an even wider and farther distribution. It is worth
mentioning that in the Chon Buri province, B. siamo­
rientalis sp. n. has been found only in Phanat Nikhom
district which is next to Chachoengsao province, while
in the eastern part of Chon Buri, that is connected to
other provinces, only B. splendens has been found.
Field notes. Betta siamorientalis sp. n. was found in
the shallow freshwater marshes, grass fields, and pad-
dy fields. It was also caught at the water edge (with
dense vegetation) of ponds, lagoons, ditches, canals,
and others water ways. Leersia hexandra (family
392 Kowasupat et al.: Betta siamorientalis, a new fighting fish from eastern Thailand

Table 1. Morphometric data of Betta siamorientalis sp. n., Betta imbellis, and Betta splendens.

Betta siamorientalis sp. n.

SPECIES Betta imbellis Betta splendens
Holotype Paratype
Number of specimens 1 40 40 40
Min. – Max. Mean S.D. (±) Min. – Max. Mean S.D. (±) Min. – Max. Mean S.D. (±)
Standard length (mm) 26.6 22.5 – 32.7 28.6 2.2 21.2 – 33.8 27.7 4.2 19.9 – 35.0 28.0 4.2
(% SL)
Total length * 135.7 126.5 – 145.5 133.3 4.4 125.1 – 142.5 133.3 3.6 123.8 – 139.1 132.2 3.8
Predorsal length 59.4 57.9 – 67.1 63.0 1.8 59.1 – 68.1 63.2 2.5 57.1 – 68.2 62.9 2.4
Postdorsal length 26.6 19.3 – 26.7 22.3 2.0 17.7 – 27.9 22.0 2.8 19.1 – 27.2 22.6 2.1
Preanal length* 34.1 35.0 – 42.7 38.8 2.2 35.0 – 43.0 38.5 2.4 32.1 – 44.5 37.4 2.8
Body depth 27.6 23.6 – 32.1 27.7 2.0 23.4 – 30.2 26.7 1.9 22.9 – 31.3 26.6 2.0
Caudal peduncle depth 19.1 14.8 – 21.8 18.4a 1.2 15.3 – 22.8 18.5 1.6 13.9 – 21.3 17.2 a 1.7
Dorsal fin base length 17.7 12.8 – 20.6 16.9 2.2 14.7 – 21.1 17.5b 1.6 12.1 – 19.3 15.8 b 1.7
Anal fin base length 64.4 57.6 – 70.8 63.8 3.2 57.1 – 68.4 61.4 2.8 53.3 – 70.1 61.9 3.9
Pectoral fin length 21.6 15.0 – 24.4 19.1 1.8 16.6 – 21.9 19.6 1.4 15.3 – 24.4 19.4 2.0
Pelvic fin length 43.1 20.2 – 58.2 38.5 8.6 26.8 – 52.2 39.3 5.9 21.2 – 55.0 35.5 8.0
Head length * 30.2 28.0 – 34.0 30.0 1.4 26.6 – 31.5 30.0 1.2 27.7 – 33.1 30.4 1.4
(% HL)
Snout length 19.8 18.6 – 30.0 23.9 2.3 18.5 – 28.4 22.9 2.1 18.1 – 28.6 23.1 2.3
Orbit diameter 28.0 23.7 – 31.6 27.7 1.9 24.4 – 31.0 27.7 1.8 23.9 – 28.9 26.5 1.4
Interorbital length* 9.3 7.8 – 14.2 11.1 1.5 8.3 – 13.5 10.7 1.3 8.6 – 13.7 10.6 1.5
Postorbital length 47.2 48.3 – 56.8 51.5 2.3 45.8 – 57.7 51.8 2.9 46.1 – 55.3 50.4 2.3
* Nonnormal distribution by using the Shapiro-Wilk test for normality (α = 0.05)
a B. siamorientalis sp. n. is significantly different from B. splendens (α = 0.05, Bonferroni-adjusted)

b B. imbellis is significantly different from B. splendens (α = 0.05, Bonferroni-adjusted)

Table 2. Meristic data of Betta siamorientalis sp. n., Betta imbellis, and Betta splendens.

Betta siamorientalis sp. n.

SPECIES Betta imbellis Betta splendens
Holotype Paratype
Number of specimens 1 40 40 40
Min. – Max. Mode Min. – Max. Mode Min. – Max. Mode
Subdorsal scales 7 5–7 6 5–8 6 5–7 6
Transverse scales 9 8–9 9 8–9 9 8–9 9
Lateral scales 30 28.5 – 31.5 30 28.5 – 31.5 30 28.5 – 32 30
Predorsal scales 23 20 – 26 23 21 – 26 23 20 – 27 24
Postdorsal scales 8 8 – 11 9 8 – 12 9 8 – 12 10
Lateral scales below dorsal-fin origin 15 14 – 16 15 14 – 16 16 14 – 17 16
Lateral scales below anal-fin origin 5 5–8 7 5–8 7 4–7 6
Anal fin rays (spines) 4 3–5 4 3–5 4 3–5 4
Anal fin rays (articulated) 23 21 – 25 23 21 – 26 22 22 – 26 23
Dorsal fin rays (spines) 1 1–2 1 1–2 1 1–2 1
Dorsal fin rays (articulated) 9 7 – 11 9 7 – 10 9 7 – 10 9
Pectoral fin rays 14 10 – 14 12 11 – 14 12 11 – 14 13
Vertebrate Zoology n 62 (3) 2012
Fig. 4. Map of eastern provinces of Thailand showing known
distribution of B. siamorientalis sp. n. (star: holotype, circle: pa­
ra­type), B. splendens (square), B. smaragdina (triangle).
Poaceae) and Eleocharis dulcis (family Cyperaceae)
were common in such habitats. The habitat character-
istics are quite similar to those of other members of B.
splendens group (see Fig. 5). Betta siamorientalis sp.
n. lives in still waters and uses water plants as shelter
to set up safe territory for building the bubble-nest,
courtship, and hatching of fry, the same preferences
as other species of the B. splendens group. Other laby-
rinth fishes cohabiting with B. siamorientalis sp. n. are
Trichopsis vittata, Trichopsis shalleri, Trichopsis pu­
mila, Trichopodus trichopterus, Anabas testudineus.
Other non-labyrinth fishes sharing the same habitats
are Lepi­docephalichthys hasselti, Pangio anguillaris,
Macro­gnathus siamensis, and Monopterus albus.
Etymology. The name siamorientalis is adopted as an
allusion to the type locality of the new species col-
lected in this study. It is derived from Siam, the former
name of Thailand, and orientalis, a Latin version for
east, the region of the country where the fish is located.
Remarks. There is only one indigenous species in
the catch sites. It should be noted that although most
specimens of B. siamorientalis sp. n. have reddish ver-
tical bars on the black opercle, the bar colour can vary,
for example, reddish to pale red, greenish-silvery or
just silvery or no colour in some populations. That B.
siamorientalis sp. n. is a new species and not a hybrid
is supported by the results from DNA barcoding using
a mitochondrial gene (cytochrome c oxidase I or COI)
and a the nuclear gene (internal transcribed spacer 1 or
393
ITS1) along with a phylogenetic tree (in preparation).
The results reveal that B. siamorientalis sp. n. is closer
to B. imbellis than B. splendens and other congeners.
Our accession numbers (www.ncbi.nlm.nih.gov) of the
COI gene and ITS1 gene of the holotype B. siamorien­
talis sp. n. are JQ818630 (COI) and JQ815715 (ITS1).
For those of the B. siamorientalis sp. n. paratypes and
of the B. imbellis and B. splendens see for example;
JQ818699 – 706 and JQ818708 – 23 for COI gene of
B. siamorientalis sp. n. and JQ818620 – 36 for ITS1
of B. siamorientalis sp. n.; JQ818776 – 81, JQ818783,
and JQ818786 – 96 for COI gene of B. imbellis; and
JQ818594 – 605, JQ818607, and JQ818610 – 19 for
ITS1 of B. imbellis; JQ818724 – 728, JQ818784 – 5,
and JQ818797 – 806 for COI gene of B. splendens;
and JQ818637 – 40, and JQ818608 – 9 for ITS1 gene
of B. splendens in gene bank website (www.ncbi.nlm.
nih.gov).
Discussion
Betta siamorientalis sp. n. shares common features
with other congeneric species in the B. splendens
group (see Witte & Schmidt (1992); Tan & Ng,
2005). However, upon close scrutiny of more than one
hundred specimens from the four provinces of east-
ern Thailand and one province in Cambodia (next to
the eastern border of Thailand), B. siamorientalis sp.
n. can be well distinguished from other known mem-
bers of the group: it has a slender body with a rather
blackish colour due to a smaller proportion of area on
the body for iridescent blue/green scales relative to
the black body background. It differs from B. imbel­
lis and B. smaragdina in having reddish bars on the
black opercle vs. green to bluish-green bars of the for-
mer and iridescent green scales on opercle of the lat-
ter. Nevertheless, it is rather difficult to differentiate
between the less acclimatized male and female mem-
bers of B. siamorientalis sp. n. due to the pale colour
of the body. Most B. siamorientalis have a light red-
dish colour on the opercle edge, which may lead some
people to mis-identify them as B. splendens. Indeed,
this new species has previously been described as B.
splendens by Lertpanich & Aranyavalai (2007), per-
haps because of the authors′ overreliance on the red
parallel bars on the opercle as a criterion while disre-
garding the body and fins which are similar to those of
B. imbellis. On the other hand, Kühne (2008) named
the so-called black imbellis from Cambodia as B. cf.
imbellis probably due to its general body shape and
fin colours similar to those of B. imbellis. Similarly,
Schäfer (2009) called his Vietnam black betta, B. cf.
imbellis. We realize that there may be others who have
394 Kowasupat et al.: Betta siamorientalis, a new fighting fish from eastern Thailand

a b

c d
Fig. 5. Habitats of B. siamorientalis sp. n. in eastern provinces of Thailand: (a) Chachoengsao (type locality) (12 Oct. 2010);
(b) Chon Buri (10 Jan. 2012), (c) Sa Kaeo (5 Oct. 2010), (d) Prachin Buri (10 Dec. 2010).

also noticed the distinct characters of this fish, includ- ferent from B. splendens. As an important step toward
ing Teerawat Utapong (personal communication) who distinguishing the black imbellis fish from the others,
has remarked that the fish from Kabin Buri district our extensive survey covering all eastern provinces of
(Prachin Buri province) could possibly be quite dif- Thailand (including those on the Thai border shared
Vertebrate Zoology n 62 (3) 2012
with Cambodia), yields morphological results which
show B. siamorientalis sp. n. to be a new species mem-
ber of the B. splendens group. Even though the mor-
phometric and meristic data seem to show only slight
differences in some of these characters (Tables 1, 2),
our statistical analysis, nevertheless, reveals a signifi-
cantly higher caudal peduncle depth in B. siamorien­
talis sp. n. when compared to that of B. splendens. The
lack of clear differences in morphometric and meristic
characters is consistent with Tan & Ng (2005) state-
ment that it is rather difficult to use meristic character-
istics and morphometric data to differentiate species
within members of the same group due to the rather
conservative nature of these characters (see also Tan
& Tan, 1996). The overall results, however, suggest
that B. siamorientalis sp. n. is more closely related to
B. imbellis than B. splendens despite the red vertical
bars on the opercle. This is further supported by our
DNA barcoding results (in preparation).
Based on our survey, the distribution of B. siam­
orientalis sp. n. in Thailand is apparently restricted
to certain areas in the upper eastern region, namely,
Chachoengsao, Chon Buri (only in areas bordering
Chachoengsao), Prachin Buri, Sa Kaeo. The fish is
also found at Banteay Meanchey, a Cambodian prov-
ince near Sa Kaeo. We have not found representatives
of this group as yet in lower eastern provinces namely
Rayong, Chantaburi, and Trat, as well as Koh Kong
of Cambodia (close to Trat province of Thailand).
Nevertheless, the bubble-nest builder B. splendens is
generally found in these latter areas. Recent reports on
the findings of the fish from Cambodia (Kühne, 2008;
Linke, 2009) and Vietnam (Schäfer, 2009) suggest
that at present the fish may be inhabiting certain re-
gions of eastern Thailand through parts of Cambodia
that connect the southern region of Vietnam. However,
B. imbellis, which is closely similar to B. siamorien­
talis sp. n., is found in the southernmost provinces of
Peninsular Thailand through to the upper part of pen-
insular Malaysia. In an attempt to explain the similari-
ties, palaeogeographical evidence is called upon here.
During the most recent ice age with lowered sea level,
the land of southern peninsular Thailand is believed to
have connected via a land mass, together with a ma-
jor river system(s), to the land of the eastern Thailand,
southern Cambodia, and Vietnam. Due to the rise of
sea level after the ice age, the land mass is currently
under water in the Gulf of Thailand thus separating
the two land areas (Dodson et al., 1995; Voris, 2000;
Sathiamurthy & Voris, 2006). This geographical evi-
dence may at least partially explain that the same or
very similar fish in the two places, separated by an
expanse of sea water since the big thaw, still retain
similarities; there are some visible differences due to
the time that has elapsed since then for the divergence
(allopatric speciation) in their new environments. In
395
addition to the bubble-nest builders above, the mouth-
brooder in eastern Thailand, B. prima (Kottelat,
1994) and the southern brooder B. pallida (Schindler
& Schmidt, 2004) in the southern peninsula also share
similar characters. It is thus plausible that B. siamori­
entalis sp. n. and B. imbellis have evolved from a com­
mon ancestor(s). However, we cannot explain the fact
that no mouth-brooders have been found in the upper
part of the Thai pennisula from Petchaburi to upper
Chumporn in spite of its present connectivity to the
more extreme southern areas (south of Chumporn pro­
vince) where mouth-brooders are common.
Regarding the phylogenetic relationship between
congeners, previous results from DNA barcoding of
bubble-nest builder bettas using cytochrome c oxidase
(COI) and 16S mitochondrial gene (Sriwattanarothai
et al., 2010) showed that the species in the B. splend­
ens group are well separated phylogenetically into at
least four main branches of B. splendens, B. imbellis,
B. mahachaiensis, and B. smaragdina. Betta stiktos′s
COI barcoding has now been done and shown to be
different and yet very close to that of one type of B.
smaragdina found in the Northeastern part of Thailand
(in preparation). Our preliminary results based on both
COI mitochondrial gene and internal transcribed spac-
er 1 (ITS1) nuclear gene point to B. siamorientalis sp.
n. as well separated phylogenetically from the B. im­
bellis, albeit closely related to the latter than B. splend­
ens. It is well distinct from B. splendens and other con-
geners. Our DNA barcoding results (in preparation)
also show that B. siamorientalis sp. n. have distinctly
different maternal lines from other congeners: of the
numerous B. siamorientalis sp. n. specimens studied
so far none has the COI sequence identical or nearly
identical to those of members of the B. splendens nor
B. imbellis (see accession number of COI and ITS 1
gene in Remarks section). The results thus indicate
that B. siamorientalis sp. n. is not a hybrid between
B. imbellis and B. splendens. Another result from our
barcoding work is that, despite opercular differences
among specimens of the B. siamorientalis sp. n., all
the above nuclear and mitochondrial DNA sequences
remain remarkably identical and they all belong to
this same species. Such opercular variation in terms of
presence or absence of red bars is also observed in the
large numbers of B. splendens studied by us.
At this juncture, we wish to address the value of the
above short DNA sequences in complementing other
characters for species identification. The COI DNA
for barcoding or other short DNA sequences as tools
for species differentiation and identification, have
proved to be useful (Hebert et al., 2003), albeit not
perfect in this role (Krishnamurthy & Francis, 2012).
Nevertheless these sequences are not just of any short
pieces of DNA but ones judiciously selected for the
purpose (Ratnasingham & Hebert, 2007). Some re-
396 Kowasupat et al.: Betta siamorientalis, a new fighting fish from eastern Thailand
cent publications (Dasmahapatra et al., 2010) may
emphasize certain deficiencies in the barcoding meth-
odology and the latter may not be the sole DNA-based
species distinguishing tool for the future (Taylor
& Harris, 2012), it is still helpful in complement-
ing morphological and other criteria. For example,
the various commonly bred and traded betta fighting
fish obtained by us in Thailand, be it as ornamentals
or fighters, show vast differences in terms of colour,
shape, fin, size, aggressiveness, etc., and yet the COI
evidence indicates that they all belong to the same B.
splendens wild type′s profile. This is consistent with
the fact that breeders have used B. splendens mainly
for selective breeding. Without the prior knowledge of
how this vast variety of ornamentals and bred fighters
have come about, these living things might be thought
to be different species; and yet they prove to be nearly
identical by DNA barcoding.
Here, we have another situation where B. siamo­
rientalis sp. n. and B. imbellis appearing not much
dif­ferent based on morphometric, meristic and other
external criteria thus making distinction between the
two difficult. We therefore have to resort to the DNA
evi­dence to further help us to decide whether the two
are the same or different. From the COI barcoding
(mito­chondrial DNA) and ITS1 (nuclear DNA) data
we are confident the two are different enough to be
called different species. (The nuclear DNA can be
derived from both maternal and parental lines, so it
can show up a hybrid, which is not found here.) At
present, B. imbellis and the black imbellis are known
to locate very far apart geographically. Without any
prior knowledge of their distant catch sites, even
experienced persons presented with the two fish for
the first time might perceive them to be identical or
nearly so. Some might even mistake the black imbel-
lis to be B. splendens. As mentioned above, there is a
situation similar to the above in that in appearance B.
stiktos and one type of B. smaragdina (type locality)
appear very similar. We suggest that these two lines
of complementary evidence above should be applied
to determine whether B. stiktos is different enough to
the nearest type locality B. smaragdina to be called a
different species just as in the case of B. imbellis and
B. siamorientalis sp. n.
At present, many people including breeders and
catchers still believe that all native fighting fish in the
aforementioned four provinces to be B. splendens.
People in the villages say that since their ancestral
times the fish has always been in this region, albeit
not used for fighting in the gambling den because as
the word imbellis implies, it is not very fierce. We
have never heard any story about southern B. imbellis
brought to the eastern part for either fighting or cross-
breeding. We know enough about the local fish fight-
ing scenes and practices not to believe that B. siamo­
rientalis sp. n. are the descendants of the discarded B.
imbellis (brought from the south) in recent past. When
the new species status of B. siamorientalis sp. n. has
been formally accepted, this knowledge will be dis-
seminated for conservation and protection of this spe-
cies.
Acknowledgements
We acknowledge Horst Linke and Jens Kühne for providing use-
ful information and comments. We thank Atison Phumchoosri,
Dr. Bunlung Nuangsaeng (Burapha University) and Dr. Adi­sorn
Monvises (Burapha University), and local breeders in Cha­cho­
engsao province, especially, Duangrat Ungklai, Somsak Tubsa,
Suthi Aungkasiri for providing information on collection locali-
ties and helping in collecting specimens from eastern Thailand.
Our thanks are also due to Teerawat Utapong for his informa-
tion on collection sites and distinct characteristics of the fighting
fish from Kabin Buri district, Prachin Buri province. This study
was financially supported by the Office of Higher Education
Commission (National University Research Grant allocated to
Mahidol University).
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